Article ID Journal Published Year Pages File Type
6133132 Journal of Virological Methods 2015 5 Pages PDF
Abstract

•A flow-FISH assay has been developed to identify and quantify B19V infected cells.•The assay provides analysis of the kinetics of B19V infection in a cell population.•The highly restricted B19V replication process in a cell population was confirmed.

Human parvovirus B19 (B19V) replication is a process highly dependent on the cellular environment, therefore methodologies allowing for analysis at single cell level could represent effective tools to understand cell-to cell differences in the replication process and to investigate cell-virus interactions. Fluorescence in situ hybridization (FISH) can be combined with flow cytometry (flow-FISH) to enable the detection of target nucleic acid sequences in thousands of individual cells in a short amount of time. In the present study, a flow-FISH assay based on the use of a digoxigenin-labeled genomic probe has been developed to discriminate B19V infected cells following in vitro infection of UT7/EpoS1 cell line and EPCs (erythroid progenitor cells) generated from peripheral blood mononuclear cells. In B19V infected UT7/EpoS1 and EPCs, viral nucleic acids were detected by the flow-FISH assay starting from 24 hpi up to 48 hpi. The method, used together with quantitative PCR techniques, can be very useful to describe the kinetics of B19V infection within a heterogeneous cell population.

Related Topics
Life Sciences Immunology and Microbiology Virology
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